Preservation of food products



Patented Apr. 24, 1951 Izloyd B. Jensen, Chicago, Illa, assignnr td-Swift & Company, Chicago, Ill.,aconporatin oil-IlliiioiS' No Drawing, Application November=10,a.1 948,i

Serial No. 59,5 09" The present invention relates; to the prepana tion and use of an antibiotic substance, and more particularly to the preparation; and use of an antimicrobial substance; of plant; origin.

The invention contemplates the utilization of an antibiotic substance naturally associated with tissues of the members of the plant. Maclum;

pomifera, commonly called bois darc and Osage orange. I have. found that. especially the heartwood and root of:- the Osage orange: tree yield an antibiotic substance of great potency as: lier'einafte'rmore. fully described;

Therefore; an objeota oi? the: present: invention; is to provide a: potent antibiotic substance of:

plant origini further ob'j ect of; the invention; is: to; provide;

am antibioticz substances which is non-toxicto: humans.

An additional object; or thainventi m 53120 p 10? vide an antibiotic su-bstanceewhich is: heat: stable; Another object-- of, the invention is to employ an antibiotic substancev having an;- origin in; a,

vegetative lant as: an; effectivev neservative for food-products. v 4

In achieving; the obiectsof. the present invenr;

tion, finely cut: or ground wood,.,bark, or fruitof.)

the Osage orange tree is treatedlwith a suitable. organic. solvent such. as acetone. The acetone, layer containing the constituents, of the, plant, tissue. soluble, therein s, parate'jd from he ac tone-iris'oluble fibrous" mass; the acetone removed therefrom and the extract taken, up in a. carrier solvent such' asiethanolf.

The following examples illustrate; preferred" embodiments of the invention, and the tests fall- 6 Claims. (01. 99-150) hereunder. illusftrate'fthe potenc of the ter'ial obtained in accordance with the; processes of theexa nples.

the Osage; orange ti'ee were finely giourid. The ground" wood waspl'ace'd in a flask and covered,-' with acetone; The flask was held at room termperat'ure for approximately 4'8 hours and shak'fi at" intervals: The acetone 1a=yer c0i1taihihg' the aceton E-SOlubl constituents 6f the- Osage diaifige: wddd was 'se'balfatd fioni the iiis'oh'lbl e' frialfifil by filtration,- and the acetone was distilled on? cohol (95- fierceiit) so that 1- ml. of the alcoholics.

extractz contained the acetone soluble materiai in 5 grams; ofi the. Osage? orange woods.

In order to evaluate the effectiveness; ofi, the

antibiotic: material in: the alcohol solution obtained in accordance withEXample 1, the fol lowing tfiStEWfiSZPBIIODIIIGdZ; I

Serial dilutions of the extract were Inadein 10: cc; broth tubes and, the; tubes inoculatedwith bacteria. Each: 1:0 ml; broth, tube contained ape-1 proximately 15,000 cellsoh aloud: poisoning strain; of, staphylbcoccua aureusg; Salmonella: typin mutium Bacillus nicer, Asnergz'llusmiger, Aspen. gil lus gldurfujs; and. Penz'cz'lliu'm notatum- The; tubes; were allowed t6 incubate V at, a temperature, of 37 C. an in p d for" a erial gr w h -iii, the e'ild 'Qf ;,Z;!5land 8daysf., In the followin table are tabulated the results, obtained; with the, aforesaid bacteria. The minus signs in saidj table ihdiriate no' bacterial growth, and the positivesign indicatei the presence. of: viable bac',-; teria atthe" incubation; periods specified:

Taiile 1 Organisms tested 1 7 Time Dilution m Difco Nutrient Broth Held Pseudu' Asper- Asper- Penidays Staph. ella monas BECIZZMS aureua tuphifluaresm'aer fig f g murium cens mg g a u a m 1 1}100 g 8 -h 1 a 1,; it l I I I I s l": 1 t a lilflnon mn-uung l 8 l 1 1-:2,o00.-.., i

. 8 I i I 1 ControlGulture: v A

1 Each 10 ml. contains approximately 75,000 cells.

The data given in the above table indicate Table H that the antibiotic efiects of the Osage orange wood extract vary in potency when tested against different organisms. Thus, the extract kills Orgamsmstested Staphylococcus in dilutions of 1 to 1,000,- Bacillus Dfl ti i 2 Sazmm new Asperil us 9 Aspetgil lus glaucus and Nutnent Broth days Staph. ella mimas Bacillus Penicillium notatum in dilutions as high as 1 to $3333 553;; 500 for limited periods of time. The bactericidal properties of the extract, lethally efiective m 1 in the dilutions indicated for the various or- 1:100 2 5 gamsms, were demonstrated by inoculating nu- 8 trient agar plates with 0.1 cc. of the aforesaid I dilutions which had been allowed to incubate for 1 5 5 days. After 3 days incubation of the inooui I I latedplates no viable bacteria were observed as 1 2 a result of the transfers. 2 i I. Example II.150 grams of the root of the g i Osage orange tree were finely ground. The 5 ground root was placed in a flask and covered f with acetone. The flask was held at room tem- Control 2 perature for approximately 48 hours and shaken g at intervals. The acetone layer containing the acetone-soluble constituents of the root was iEach 10 contains approximately 75,000 cells, separated from the insoluble material by filtration, and the acetone removed from the extract The data given in the above table indicate that by distillation under reduced pressure at 42 C. the antibiotic effects of the Osage orange root The residue was taken up in ethyl alcohol (95 extract vary in potency when tested against difper cent) so that 1 ml. of the alcoholic extract ferent organisms, and possesses antibiotic propcontained the acetone-soluble material in 5 grams erties somewhat different than the extract from of Osage orange root. the heartwood of the Osage orange; Thus, the

In order to evaluate the effectiveness of the root extract kills Staphylococcus aureus and Baantibiotic material in the alcohol solution obcillas niger in dilutions of 1 to 1,000, but has tained in accordance with Example II, the follittle effect on the other organisms tested. The lowing test was performed: bactericidal properties of the extract were dem- Serial dilutions of the extract were made in onstrated in the same manner as in Example I. 10 cc. broth tubes and the tubes inoculated with Although acetone is the preferred primary solbacteria. Each 10ml. broth tube contained apvent, I have found that other organic solvents proximately 75,000 cells of afood poisoning strain 40 are suitable as the extracting solvent. For exof Staphylococcus aureus, Salmonella typhimuample, dioxane, ethyl acetate, methyl ethyl kerium, Bacillus m'ger, Aspergillus niger, Aspergillus tone and ethanol may be used. I have also found glaucus and Penicillium notatum. The tubes a water extraction of the antibiotic material were allowed to incubate at a temperature of to be only slightly effective as an antibiotic be- 37 C. and inspected for bacterial growth at the cause of the proteinaceous material dissolved end of 1, 2, 5 and 8 days. In the following table therein. are tabulated the results obtained with the afore- The heat stability of the antibiotic material said bacteria. The minus signs in said table inw er i y h in a p r f h i dicate no bacterial growth, and the positive signs material and then inoculating with B. nz'ger, a indicate the presence of viable bacteria at the invariant strain of B. subtz'lis. The results of the cubation periods specified. foregoing tests are set forth in Table III.

Table III t: In b Dilutions of Heated antibiotic g g Temp. stilt days 200 500 1,000 2,000 5,000 C. 1 15 4 i iii I 1% 4 rai iiii 3% 4 a 6 nastiest It is obvious=that many variations in the solvc'fitvtreatingprocess may-be advantageously; em-

ployed. For example, since the antibioticmate-- ture of solvents; Ihave found itpreferable to process the antibiotic substance contained in the osageorangeby a two=-stage" solvent process" involviirg first," treating" the" substance' with a" pl'i'i' mary solvent which is substantially removed'and then taking up the resulting" extract" with" a ctiierscitent so wforma solution which is" aconvenient form of using the producti Ihave" term-d4 that acetone is" very effective primary,

servers in producing the antibiotic: substance from the plantti's'sue'f Afterevapo'ratingofithe" acetone, the preferred solution is made with a carrier solvent such as ethanol, which may be consumed internally without harm. By using ethyl alcohol I also utilize its well-known properties, such as non-toxicity and ability to prevent contamination.

In the processing of food products a serious problem is the preservation thereof, and although great strides have been made in improving the quality and keeping time of various food products by refrigeration, canning, curing, plant sanitation, etc., the problem of preservation of food requires the constant attention of the food processor to improve long-established methods of treating food and to devise new means of food control.

Food spoilage is due primarily to microbial action, and the species of microorganisms responsible for food deterioration are numerous. The sporing genera, Clostridium and Bacillus, for example, when contaminatin food, cause food spoilage. Another serious concern of the food handler is food poisoning, which is often of an insidious nature because food products may be contaminated with one or more of the harmful bacteria referred to herein and. yet appear organoleptically sound. Among the food poisoning bacteria, the most comm-on food poisoner is Staphylococcus.

The antibacterial substance of the present invention may be used to retard or prevent microbial growth in various food products, all of which tend to harbor pathogenic organisms in varying degrees. in, therefore, may advantageously be employed, for example, in treating meat products, such as hams, sausages and canned meats; filled pastries, such as cream puffs, chocolate eclairs, custards, and the like; and canned vegetables to prevent swelling due to organisms, such as the genus Bacillus.

The invention will be more fully understood from the following description of methods of using the antibiotic substance.

As hereinbefore mentioned, I have found that the antibiotic reagent herein described is lethal to a number of microbes in a 1 to 1,000 dilution of the extract, the extract being of such strength that 1 ml. of the alcoholic solution thereof contained the acetone-soluble constituents in 5 grams of root. Therefore, the addition of 1 part of the solution to 1,000 parts of the conventional pickling solution will result in a meat product cured The antibiotic material disclosed herea. therewith: which has" a ver' *"marlied iii-crease in resistance to the development of pathogenic organisms g The following isfan ex'ample' of a-curihg pickle stock; W'l iich maylo'e-used in arprocess for quick curing Hams when provided withantibiotic' substance i'rr'theproportions outlihedaboveqthat is; the antibiotic substance is'addedirr an amountsuch as toeffect a 1 to 1,000 dili1tion'of"tlfeanti metre materiar of the ar itrary strengtli herein described (*lfml-E v of-rootfi V To l'lm gallon's of a li salometersoluticwthere are added:

'l lbs sodium-nitrate:

10:02: sodium'fiitrite's 378:5 cc ..antibiotic'solutiom Luncheon meat was? cured. according. to. con, ventional methods in which the antibiotic substancemas-presentima 1 to 1000 dilution. The meat product was placed in six 6-pound cans and retorted to an inside temperature of 155 F. and incubated at 99 F. After 30 days of incubation no swelling of the cans could be observed. The control cans swelled in one week.

A further example of the utility of the antibiotic material is their use in a cream filling for various pastries. To determine the effectiveness, a standard cream filling used in eclairs was inoculated with a bacterial solution containing 100,000 living Staphylococcus aureus S-l per 100 ml. and the filling incubated at a temperature between 75 F. and F. The results of the tests are given in Table IV.

Table IV Incubated at 75 F. Dilution sltaaghylococcus (S-l) per Antibiotic in cream g filling 24 hrs. 48 hrs. 72 hrs.

Osage orange wood l-300 2,000, 000 N o antibiotic 0 140, 000, 000 No staphylococci 0 0 0 0 The foregoing tests indicate that the antibiotic preparation exerts a pronounced inhibiting action against the Staphylococcus aureus in cream fillings.

Obviously, many modifications and variations of the invention, as hereinbefore set forth, may be made without departing from the spirit and scope thereof, and therefore only such limitations should be imposed as are indicated in the appended claims.

I claim:-

1. In the process of treating food products, the step of adding to a food product a small amount of an alcoholic solution of antibiotic substance derived from the plant Maclum pomz'fera to preserve said food product against pathogenic bacteria responsible for food poisoning and spoilage.

2. In the process of quick-curing meats, wherein a pickling solution is injected into said meat, the step of adding to said pickling solution a small amount of the antibiotic substance derived from the plant Maclura pomifera by treatment with a non-aqueous organic solvent whereby the growth of pathogenic bacteria responsible for food spoilage and poisoning in the meat is substantially retarded.

3. In the process of preparing pastries containing fillings subject to spoilage by pathogenic bacteria, the step of adding to the said filling a small amount of the antibiotic substance derived from the plant M aclum pomifera by treatment with a non-aqueous organic solvent whereby the growth of pathogenic bacteria issubstantially retarded.

4. A meat pickle including a small amount of an antibiotic substance derived from the plant M aclura pomifera by treatment with a nonaqueous organic solvent.

5.- A cream filling for pastries including a small amount of an antibiotic substance derived from the plant M aclura pomz'fera by treatment with a non-aqueous organic solvent.

6. In the process of treating food products, the step of adding to a food product a smallamount of an antibiotic substance derived from the plant Maclura pomifera by treatment with a nonaqueous organic solvent to preserve said food product against pathogenic bacteria responsible for food poisoning and spoilage.

LLOYD B. JENSEN.

REFERENCES CITED The following references are of record in the file of this patent:

UNITED STATES PATENTS Number Name Date 469,850 Chesebrough Mar. 1, 1892 2,084,864 Paddock June 22, 1937 2,098,110 Schertz et a1 Nov. 2, 1937 2,117,478 Hall May 17, 1938 2,180,750 Urbain Nov. 21, 1939 OTHER REFERENCES Nature, May 13, 1944, vol. 153, page 598, article entitled Antibacterial Substances in Green Plants.

Chemical and Engineering News, September 1945, page 1622, article entitled Penicillin as a Preservative.

The American Womans Cook Book, 1945, by Ruth Berolzheimer, published by Consolidated Book Publishers, Chicago, page 475. 

1. IN THE PROCESS OF TREATING FOOD PRODUCTS, THE STEP OF ADDING TO A FOOD PRODUCT A SMALL AMOUNT OF AN ALCOHOLIC SOLUTION OF ANTIBIOTIC SUBTANCE DERIVED FROM THE PLANT MACLURA POMIFERA TO PRESERVE SAID FOOD PRODUCT AGAINST PATHOGENIC BACTERIA RESPONSIBLE FOR FOOD POISONING AND SPOILAGE. 